Involvement of Catalase and Superoxide Dismutase in Resistance ofBotrytis cinereato Dicarboximide Fungicide Vinclozolin

Abstract In order to examine whether defensive enzymes are related to the resistance of Botrytis cinerea to dicarboximide fungicides, catalase (CAT) and superoxide dismutase (SOD) activities were compared in a dicarboximide-susceptible (DS) isolate and a dicarboximide-resistant (DR) isolate of B. cinerea . In the absence of vinclozolin, the DR isolate had 1.5- and 3.0-fold higher CAT and SOD activities, respectively, than the DS isolate. When the DR isolate was incubated with 2 μM vinclozolin, CAT and SOD activities were considerably enhanced at 1 h after the fungicide treatment and then decreased to the level of their respective control. In the DS isolate, however, such changes of the enzyme activities were not significant. Native polyacrylamide gel electrophoresis and activity staining of CAT and SOD revealed that both enzymes had two isoforms and that the staining intensities of the isoforms were different in the DS and the DR isolates. Both isoforms of CAT appeared to have higher staining intensity in the DR isolate than in the DS isolate, whereas one isoform of SOD was more preferentially distinct in the DR isolate. The enhanced isoform of SOD in the DR isolate was found to be Cu,Zn-SOD. When the DR isolate was incubated with 2 μM vinclozolin, both isoforms of CAT from the DR isolate tended to become slightly more apparent at 1 h after the fungicide treatment, but their staining activity was slightly reduced after the prolonged treatment of the fungicide. No such changes of the staining intensity were observed in the CAT isoforms from the DS isolate and in the SOD isoforms either from the DS or from the DR isolate. Our results demonstrate that the DR isolate has enhanced activities of CAT and SOD regardless of the presence of vinclozolin. They also show that a much higher amount of the Cu,Zn-SOD isoform exists in the DR isolate than in the DS isolate. Therefore, it can be concluded that enhanced activities of CAT and SOD, mostly of Cu,Zn-SOD, at least partly confer the resistance of B. cinerea to the dicarboximide fungicides.

[1]  C. Steel,et al.  Catalase activity and sensitivity to the fungicides, iprodione and fludioxonil in Botrytis cinerea , 1996 .

[2]  G. Choi,et al.  Lipid Peroxidation and Membrane Disruption by Vinclozolin in Dicarboximide-Susceptible and -Resistant Isolates ofBotrytis cinerea , 1996 .

[3]  G. Paliyath,et al.  Ultraviolet-B- and Ozone-Induced Biochemical Changes in Antioxidant Enzymes of Arabidopsis thaliana , 1996, Plant physiology.

[4]  K.,et al.  Changes in Isozyme Profiles of Catalase, Peroxidase, and Glutathione Reductase during Acclimation to Chilling in Mesocotyls of Maize Seedlings , 1995, Plant physiology.

[5]  Sami Ahmad.  Oxidative Stress and Antioxidant Defenses in Biology , 1995, Springer US.

[6]  W. Edlich.  Mechanism of action of dicarboximide fungicides , 1995 .

[7]  Sami Ahmad.  Antioxidant Mechanisms of Enzymes and Proteins , 1995 .

[8]  H. Lyr,et al.  Modern selective fungicides : properties, applications, mechanisms of action , 1987 .

[9]  D. A. Dalton.  Antioxidant Defenses of Plants and Fungi , 1995 .

[10]  Christine H. Foyer,et al.  Photooxidative stress in plants , 1994 .

[11]  C. Steel,et al.  The Physiological Basis of Resistance to the Dicarboximide Fungicide Iprodione in Botrytis cinerea , 1993 .

[12]  J. G. Scandalios.  Oxygen Stress and Superoxide Dismutases , 1993, Plant physiology.

[13]  H. Lyr,et al.  Target sites of fungicides with primary effects on lipid peroxidation , 1992 .

[14]  M. Tien,et al.  An investigation into the role of lipid peroxidation in the mode of action of aromatic hydrocarbon and dicarboximide fungicides , 1992 .

[15]  Dirk Inzé,et al.  SUPEROXIDE DISMUTASE AND STRESS TOLERANCE , 1992 .

[16]  W. Koeller.  Target Sites of Fungicide Action , 1991 .

[17]  J. P. Spychalla,et al.  Superoxide dismutase, catalase, and α-tocopherol content of stored potato tubers , 1990 .

[18]  R. Pardini,et al.  Mechanisms for regulating oxygen toxicity in phytophagous insects. , 1990, Free radical biology & medicine.

[19]  B. Halliwell,et al.  Role of free radicals and catalytic metal ions in human disease: an overview. , 1990, Methods in enzymology.

[20]  R. Beever,et al.  Strains of Botrytis cinerea resistant to dicarboximide and benzimidazole fungicides in New Zealand vineyards , 1989 .

[21]  E. Cadenas,et al.  Biochemistry of oxygen toxicity. , 1989, Annual review of biochemistry.

[22]  K. Brent.  Modern selective fungicides—properties, applications and mechanisms of action: ed. by H. Lyr. ISBN 0 582 00461 6 (383 pp; £35.00). Harlow and New York: Longmans. 1987 , 1988 .

[23]  H. Lyr.  Lipid peroxidation: A side effect of sterol demethylation inhibitor fungicides in Mucor mucedo (L.) fres and Ustilago maydis , 1988 .

[24]  M. Wolfe,et al.  Populations of Plant Pathogens: Their Dynamics and Genetics , 1987 .

[25]  G. Matters,et al.  Effect of the free radical-generating herbicide paraquat on the expression of the superoxide dismutase (Sod) genes in maize. , 1986, Biochimica et biophysica acta.

[26]  M. Duke,et al.  Purification and characterization of an iron-containing superoxide dismutase from a eucaryote, Ginkgo biloba. , 1985, Archives of biochemistry and biophysics.

[27]  J. F. Ryley,et al.  Mode of action of antifungal agents , 1984 .

[28]  H. Aebi,et al.  Catalase in vitro. , 1984, Methods in enzymology.

[29]  B. K. Cooke,et al.  Insensitivity of Botrytis cinerea to iprodione, procymidone and vinclozolin and their uptake by the fungus , 1979 .

[30]  J. G. Scandalios,et al.  Developmental Expression and Intracellular Localization of Superoxide Dismutases in Maize , 1979 .

[31]  I. Fridovich,et al.  Superoxide Dismutase and Oxygen Metabolism in Streptococcus faecalis and Comparisons with Other Organisms , 1978, Journal of bacteriology.

[32]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[33]  K. Asada,et al.  Superoxide dismutases from a blue-green alga, Plectonema boryanum. , 1975, The Journal of biological chemistry.

[34]  R. Nalbandyan,et al.  Interaction of hydrogen peroxide with superoxide dismutase from erythrocytes , 1972, FEBS letters.

[35]  I. Fridovich,et al.  The purification and properties of superoxide dismutase from Neurospora crassa. , 1972, The Journal of biological chemistry.

[36]  G. Rotilio,et al.  A pulse radiolysis study of superoxide dismutase. , 1972, Biochimica et biophysica acta.

[37]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[38]  B. Little,et al.  Xanthine oxidase for calibration of the oxygen electrode apparatus. , 1970, Analytical biochemistry.